9,233 Matching Annotations
  1. Mar 2021
    1. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 4.7

      AssayResultAssertion: Normal

      StandardErrorMean: 0.1

    2. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 4.7

      AssayResultAssertion: Normal

      StandardErrorMean: 0.92

    3. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 4.7

      AssayResultAssertion: Normal

      StandardErrorMean: 0.06

    4. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 4.7

      AssayResultAssertion: Normal

      StandardErrorMean: 0.11

    5. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 4.8

      AssayResultAssertion: Normal

      StandardErrorMean: 0.23

    6. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 4.8

      AssayResultAssertion: Normal

      StandardErrorMean: 0.79

    7. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 4.8

      AssayResultAssertion: Normal

      StandardErrorMean: 0.24

    8. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 4.8

      AssayResultAssertion: Normal

      StandardErrorMean: 0.67

    9. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 4.9

      AssayResultAssertion: Normal

      StandardErrorMean: 0.29

    10. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 4.9

      AssayResultAssertion: Normal

      StandardErrorMean: 0.93

    11. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 4.9

      AssayResultAssertion: Normal

      StandardErrorMean: 0.44

    12. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 4.9

      AssayResultAssertion: Normal

      StandardErrorMean: 0.71

    13. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 4.9

      AssayResultAssertion: Normal

      StandardErrorMean: 0.01

    14. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 4.9

      AssayResultAssertion: Normal

      StandardErrorMean: 0.08

    15. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 4.9

      AssayResultAssertion: Normal

      StandardErrorMean: 0.04

    16. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 4.9

      AssayResultAssertion: Normal

      StandardErrorMean: 0.49

    17. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 4.9

      AssayResultAssertion: Normal

      StandardErrorMean: 0.57

    18. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 4.9

      AssayResultAssertion: Normal

      StandardErrorMean: 0.27

    19. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 4.9

      AssayResultAssertion: Normal

      StandardErrorMean: 0.32

    20. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 4.9

      AssayResultAssertion: Normal

      StandardErrorMean: 0.1

    21. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 5

      AssayResultAssertion: Normal

      StandardErrorMean: 0.6

    22. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 5

      AssayResultAssertion: Normal

      StandardErrorMean: 0.08

    23. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 5

      AssayResultAssertion: Normal

      StandardErrorMean: 0.15

    24. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 5

      AssayResultAssertion: Normal

      StandardErrorMean: 1.01

    25. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 5

      AssayResultAssertion: Normal

      StandardErrorMean: 0.58

    26. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 5

      AssayResultAssertion: Normal

      StandardErrorMean: 0.32

    27. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 5

      AssayResultAssertion: Normal

      StandardErrorMean: 1.21

    28. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 5.1

      AssayResultAssertion: Normal

      StandardErrorMean: 0.56

    29. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 5.1

      AssayResultAssertion: Normal

      StandardErrorMean: 0.7

    30. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 5.1

      AssayResultAssertion: Normal

      StandardErrorMean: 1

    31. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 5.2

      AssayResultAssertion: Normal

      StandardErrorMean: 0.39

    32. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 5.2

      AssayResultAssertion: Normal

      StandardErrorMean: 0.1

    33. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 5.2

      AssayResultAssertion: Normal

      StandardErrorMean: 0.33

    34. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 5.2

      AssayResultAssertion: Normal

      StandardErrorMean: 0.7

    35. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 5.3

      AssayResultAssertion: Normal

      StandardErrorMean: 0.84

    36. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 5.3

      AssayResultAssertion: Normal

      StandardErrorMean: 0.46

    37. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 5.4

      AssayResultAssertion: Normal

      StandardErrorMean: 0.13

    38. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 5.6

      AssayResultAssertion: Normal

      StandardErrorMean: 0.16

    39. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 5.8

      AssayResultAssertion: Normal

      StandardErrorMean: 1.15

    40. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 5.8

      AssayResultAssertion: Normal

      StandardErrorMean: 0.1

    41. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 6

      AssayResultAssertion: Normal

      StandardErrorMean: 0.13

    42. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 6.2

      AssayResultAssertion: Normal

      StandardErrorMean: 1.39

    43. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 6.4

      AssayResultAssertion: Normal

      StandardErrorMean: 0.45

    44. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 6.4

      AssayResultAssertion: Normal

      StandardErrorMean: 0.96

    45. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 6.5

      AssayResultAssertion: Normal

      StandardErrorMean: 1.55

    46. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 6.6

      AssayResultAssertion: Normal

      StandardErrorMean: 0.05

    47. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 6.6

      AssayResultAssertion: Normal

      StandardErrorMean: 0.8

    48. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 6.9

      AssayResultAssertion: Normal

      StandardErrorMean: 0.35

    49. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 7.1

      AssayResultAssertion: Normal

      StandardErrorMean: 0.43

    50. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 7.2

      AssayResultAssertion: Normal

      StandardErrorMean: 0.01

    51. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 7.3

      AssayResultAssertion: Normal

      StandardErrorMean: 0.08

    52. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 7.5

      AssayResultAssertion: Normal

      StandardErrorMean: 0.61

    53. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 7.7

      AssayResultAssertion: Normal

      StandardErrorMean: 0.15

    54. Results for individual PALB2 variants were normalized relative to WT-PALB2 and the p.Tyr551ter (p.Y551X) truncating variant on a 1:5 scale with the fold change in GFP-positive cells for WT set at 5.0 and fold change GFP-positive cells for p.Y551X set at 1.0. The p.L24S (c.71T>C), p.L35P (c.104T>C), p.I944N (c.2831T>A), and p.L1070P (c.3209T>C) variants and all protein-truncating frame-shift and deletion variants tested were deficient in HDR activity, with normalized fold change <2.0 (approximately 40% activity) (Fig. 1a).

      AssayResult: 7.7

      AssayResultAssertion: Normal

      StandardErrorMean: 0.15

    55. A total of 84 PALB2 patient-derived missense variants reported in ClinVar, COSMIC, and the PALB2 LOVD database were selected

      HGVS: NM_024675.3:c.100C>T p.(Arg34Cys)

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    1. Olaparib sensitivity assayFor the sensitivity assay in HeLa, 240 000 cells were seeded into one well of a six-well plate before being transfected 6–8 h later with 50 nM control or PALB2 siRNA using Lipofectamine RNAiMAX (Invitrogen). The next morning, cells were complemented with 800 ng of the peYFP-C1 empty vector or the indicated siRNA-resistant YFP-tagged PALB2 construct using Lipofectamine 2000 (Invitrogen) for 24 h and then seeded in triplicates into a Corning 3603 black-sided clear bottom 96-well microplate at a density of 3000 cells per well. The remaining cells were kept and stored at −80°C until processed for protein extraction and immunoblotting as described above. Once attached to the plate, cells were exposed to different concentrations of olaparib (Selleckchem, #S1060) ranging from 0 (DMSO) to 2.5 μM. After 3 days of treatment, nuclei were stained with Hoechst 33342 (Invitrogen) at 10 μg/ml in media for 45 min at 37°C. Images of entire wells were acquired at 4x with a Cytation 5 Cell Imaging Multi-Mode Reader followed by quantification of Hoechst-stained nuclei with the Gen5 Data Analysis Software v3.03 (BioTek Instruments). Cell viability was expressed as percentage of survival in olaparib-treated cells relative to vehicle (DMSO)-treated cells. Results represent the mean ± SD of at least 3 independent experiments, each performed in triplicate.

      AssayGeneralClass: BAO:0003009 cell viability assay

      AssayMaterialUsed: CLO:0003684 HeLa cell

      AssayDescription: HeLa cells were treated with PALB2 siRNA followed by transfection peYFP-PALB2 expressing PALB2 variants (or empty vector) and exposed to olaparib (2.5 µM) for 3 days. Nuclei were stained with Hoechst 33342 and measured as an indicator of cell viability.

      AssayReadOutDescription: Cell viability expressed as percentage of survival in olaparib-treated cells relative to vehicle (DMSO)-treated cells

      AssayRange: %

      AssayNormalRange: Not reported

      AssayAbnormalRange: Not reported

      AssayIndeterminateRange: Not reported

      ValidationControlPathogenic: 1

      ValidationControlBenign: 3

      Replication: At least 3 independent experiments, each performed in triplicate

      StatisticalAnalysisDescription: Kruskal–Wallis test with Dunn's multiple comparison post-test

    2. SUPPLEMENTARY DATA

      AssayResult: 5

      AssayResultAssertion: Abnormal

      PValue: < 0.0001

      Approximation: Exact assay result value not reported; value estimated from Figure 6C.

    3. SUPPLEMENTARY DATA

      AssayResult: -98

      AssayResultAssertion: Abnormal

      PValue: < 0.0001

    4. SUPPLEMENTARY DATA

      AssayResult: 48

      AssayResultAssertion: Abnormal

      PValue: < 0.0001

      Approximation: Exact assay result value not reported; value estimated from Figure 1D.

      ControlType: Abnormal; empty vector

    5. SUPPLEMENTARY DATA

      AssayResult: 100

      AssayResultAssertion: Normal

      ControlType: Normal; wild type PALB2 cDNA

    6. SUPPLEMENTARY DATA

      AssayResult: 106

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    7. SUPPLEMENTARY DATA

      AssayResult: 108.6

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    8. SUPPLEMENTARY DATA

      AssayResult: 64.45

      AssayResultAssertion: Abnormal

      PValue: < 0.0001

      Comment: Exact values reported in Table S3.

    9. SUPPLEMENTARY DATA

      AssayResult: 84.49

      AssayResultAssertion: Indeterminate

      PValue: 0.0058

      Comment: Exact values reported in Table S3.

    10. SUPPLEMENTARY DATA

      AssayResult: 92.43

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    11. SUPPLEMENTARY DATA

      AssayResult: 88.66

      AssayResultAssertion: Not reported

      PValue: 0.727

      Comment: Exact values reported in Table S3.

    12. SUPPLEMENTARY DATA

      AssayResult: 96.63

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    13. SUPPLEMENTARY DATA

      AssayResult: 97.59

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    14. SUPPLEMENTARY DATA

      AssayResult: 94.36

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    15. SUPPLEMENTARY DATA

      AssayResult: 98.94

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    16. SUPPLEMENTARY DATA

      AssayResult: 87.19

      AssayResultAssertion: Not reported

      PValue: 0.341

      Comment: Exact values reported in Table S3.

    17. SUPPLEMENTARY DATA

      AssayResult: 98.25

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    18. SUPPLEMENTARY DATA

      AssayResult: 57.61

      AssayResultAssertion: Abnormal

      PValue: < 0.0001

      Comment: Exact values reported in Table S3.

    19. SUPPLEMENTARY DATA

      AssayResult: 109.2

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    20. SUPPLEMENTARY DATA

      AssayResult: 95.47

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    21. SUPPLEMENTARY DATA

      AssayResult: 97.77

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    22. SUPPLEMENTARY DATA

      AssayResult: 103.5

      AssayResultAssertion: Normal

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    23. SUPPLEMENTARY DATA

      AssayResult: 100.7

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    24. SUPPLEMENTARY DATA

      AssayResult: 102.6

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    25. SUPPLEMENTARY DATA

      AssayResult: 77.32

      AssayResultAssertion: Indeterminate

      PValue: 0.0002

      Comment: Exact values reported in Table S3.

    26. SUPPLEMENTARY DATA

      AssayResult: 82.22

      AssayResultAssertion: Indeterminate

      PValue: 0.004

      Comment: Exact values reported in Table S3.

    27. SUPPLEMENTARY DATA

      AssayResult: 96.97

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    28. SUPPLEMENTARY DATA

      AssayResult: 102.1

      AssayResultAssertion: Normal

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    29. SUPPLEMENTARY DATA

      AssayResult: 101.6

      AssayResultAssertion: Normal

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    30. SUPPLEMENTARY DATA

      AssayResult: 109.7

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    31. SUPPLEMENTARY DATA

      AssayResult: 109.4

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    32. SUPPLEMENTARY DATA

      AssayResult: 107.5

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    33. SUPPLEMENTARY DATA

      AssayResult: 100.5

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    34. SUPPLEMENTARY DATA

      AssayResult: 103.3

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    35. SUPPLEMENTARY DATA

      AssayResult: 108.7

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    36. SUPPLEMENTARY DATA

      AssayResult: 106.8

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    37. SUPPLEMENTARY DATA

      AssayResult: 94.01

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    38. SUPPLEMENTARY DATA

      AssayResult: 92.68

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    39. SUPPLEMENTARY DATA

      AssayResult: 92.03

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    40. SUPPLEMENTARY DATA

      AssayResult: 93.06

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    41. SUPPLEMENTARY DATA

      AssayResult: 86.49

      AssayResultAssertion: Not reported

      PValue: 0.3376

      Comment: Exact values reported in Table S3.

    42. SUPPLEMENTARY DATA

      AssayResult: 76.21

      AssayResultAssertion: Indeterminate

      PValue: 0.0001

      Comment: Exact values reported in Table S3.

    43. SUPPLEMENTARY DATA

      AssayResult: 85.76

      AssayResultAssertion: Indeterminate

      PValue: 0.0445

      Comment: Exact values reported in Table S3.

    44. SUPPLEMENTARY DATA

      AssayResult: 47.64

      AssayResultAssertion: Abnormal

      PValue: < 0.0001

      Comment: Exact values reported in Table S3.

    45. SUPPLEMENTARY DATA

      AssayResult: 86.51

      AssayResultAssertion: Not reported

      PValue: 0.2166

      Comment: Exact values reported in Table S3.

    46. SUPPLEMENTARY DATA

      AssayResult: 97.46

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    47. SUPPLEMENTARY DATA

      AssayResult: 91.53

      AssayResultAssertion: Not reported

      PValue: > 0.9999

      Comment: Exact values reported in Table S3.

    48. SUPPLEMENTARY DATA

      AssayResult: 82.06

      AssayResultAssertion: Indeterminate

      PValue: 0.0058

      Comment: Exact values reported in Table S3.

    49. SUPPLEMENTARY DATA

      AssayResult: 76.45

      AssayResultAssertion: Indeterminate

      PValue: 0.0001

      Comment: Exact values reported in Table S3.

    50. SUPPLEMENTARY DATA

      AssayResult: 86.74

      AssayResultAssertion: Not reported

      PValue: 0.1836

      Comment: Exact values reported in Table S3.

    51. To this end, 44 missense variants found in breast cancer patients were identified in the ClinVar database (https://www.ncbi.nlm.nih.gov/clinvar) and/or selected by literature curation based on their frequency of description or amino acid substitution position in the protein (Supplemental Table S1).

      HGVS: NM_024675.3:c.104T>C p.(Leu35Pro)

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    1. Reviewer #1 (Public Review):

      The authors used a CRISPR screen to investigate the basis of metastasis of ovarian cancer (OC) cells. Overall, they identified two key genes, IL20RA, one of which was studied in detail. They identify an IL20/IL20RA communication between ovarian cancer cells and peritoneal mesothelial cells to promote M1 macrophages and prevent dissemination of the cancer cells. IL-20 mediated crosstalk is blocked in metastasized OC cells by decreased expression of IL-20RA. Interestingly, IL20RA is also decreased in cells from OC patients with peritoneal metastasis, and reconstitution of IL20RA in metastatic OC cells suppresses metastasis. Moreover, OC cells induce mesothelial cells to produce IL20 and IL24.

      Overall, this is a nice study. It is well-written, and the data are clear. A range of methodologies are used that support the conclusions, with both over-expression and under-expression related studies supporting some key conclusions.

      The overall model is that there is crosstalk between disseminated OC cells and mesothelial cells and macrophages. OC cells when disseminated into the peritoneal cavity stimulate mesothelial cells to produced IL20 and IL24, which via IL20RA trigger STAT3 to produced OAS/RNase L and production of IL-18, to promote an M1 phenotype. The M1 phenotype lowers metastasis. Highly metastatic cells block this pathway by decreasing IL20RA expression.

      These findings are interesting, with potential therapeutic ramifications.

    1. A cell-based functional assay for PALB2 variants

      AssayGeneralClass: BAO:0003061 reporter protein

      AssayMaterialUsed: CLO:0037317 mouse embryonic stem cell line

      AssayDescription: Stable expression of wild type and variant PALB2 cDNA constructs in Trp53 and Palb2-null mouse cell line containing DR-GFP reporter; I-SceI endonuclease introduces a double-stranded break in the reporter construct and efficient repair results in GFP expression, which is detected by flow cytometry

      AssayReadOutDescription: Relative homologous recombination (HR) efficiency represented as mean percentages of GFP-positive cells among the mCherry-positive cells relative to wild type, which was set to 100%

      AssayRange: %

      AssayNormalRange: HR levels comparable to that of cells expressing wild type PALB2; no numeric threshold given

      AssayAbnormalRange: HR levels ≤40% of wild type

      AssayIndeterminateRange: Not reported

      ValidationControlPathogenic: 12

      ValidationControlBenign: 9

      Replication: 2 independent experiments

      StatisticalAnalysisDescription: Not reported

    2. Source Data

      AssayResult: 115.71

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardErrorMean: 3.09

      Comment: Exact values reported in “Source Data” file.

    3. Source Data

      AssayResult: 80.95

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 0.01

      StandardErrorMean: 0

      Comment: Exact values reported in “Source Data” file. Discrepancy in “Supplementary Data 1” file: nucleotide reported as c.3191A>G.

    4. Source Data

      AssayResult: 101.02

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 5.45

      StandardErrorMean: 3.86

      Comment: Exact values reported in “Source Data” file. Discrepancy in “Source Data” file: protein reported as T1064C.

    5. Source Data

      AssayResult: 84.43

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 2.77

      StandardErrorMean: 1.96

      Comment: Exact values reported in “Source Data” file. Discrepancy in “Source Data” file: protein reported as L855P (based on matching values reported in the “Supplementary Data 1” file to values reported in the “Source Data” file.

    6. Source Data

      AssayResult: 15.58

      AssayResultAssertion: Abnormal

      ReplicateCount: 6

      StandardDeviation: 0.52

      StandardErrorMean: 0.37

      ControlType: Abnormal; empty vector (set 5)

      Comment: Exact values reported in “Source Data” file.

    7. Source Data

      AssayResult: 7.93

      AssayResultAssertion: Abnormal

      ReplicateCount: 6

      StandardDeviation: 0.56

      StandardErrorMean: 0.39

      ControlType: Abnormal; empty vector (set 4)

      Comment: Exact values reported in “Source Data” file.

    8. Source Data

      AssayResult: 8.71

      AssayResultAssertion: Abnormal

      ReplicateCount: 6

      StandardDeviation: 1.75

      StandardErrorMean: 1.24

      ControlType: Abnormal; empty vector (set 3)

      Comment: Exact values reported in “Source Data” file.

    9. Source Data

      AssayResult: 7.11

      AssayResultAssertion: Abnormal

      ReplicateCount: 6

      StandardDeviation: 2.37

      StandardErrorMean: 1.68

      ControlType: Abnormal; empty vector (set 2)

      Comment: Exact values reported in “Source Data” file.

    10. Source Data

      AssayResult: 7.83

      AssayResultAssertion: Abnormal

      ReplicateCount: 6

      StandardDeviation: 1.13

      StandardErrorMean: 0.8

      ControlType: Abnormal; empty vector (set 1)

      Comment: Exact values reported in “Source Data” file.

    11. Source Data

      AssayResult: 100

      AssayResultAssertion: Normal

      ReplicateCount: 38

      StandardDeviation: 0

      StandardErrorMean: 0

      ControlType: Normal; wild type PALB2 cDNA

      Comment: Exact values reported in “Source Data” file.

    12. Source Data

      AssayResult: 97.16

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 1.32

      StandardErrorMean: 0.93

      Comment: Exact values reported in “Source Data” file.

    13. Source Data

      AssayResult: 30.35

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 3.64

      StandardErrorMean: 2.57

      Comment: Exact values reported in “Source Data” file.

    14. Source Data

      AssayResult: 20.32

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 0.49

      StandardErrorMean: 0.35

      Comment: Exact values reported in “Source Data” file.

    15. Source Data

      AssayResult: 7.42

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 2.87

      StandardErrorMean: 2.03

      Comment: Exact values reported in “Source Data” file.

    16. Source Data

      AssayResult: 91.47

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 8.63

      StandardErrorMean: 6.1

      Comment: Exact values reported in “Source Data” file.

    17. Source Data

      AssayResult: 100.19

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 5.64

      StandardErrorMean: 3.99

      Comment: Exact values reported in “Source Data” file.

    18. Source Data

      AssayResult: 10.53

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 2.79

      StandardErrorMean: 1.97

      Comment: Exact values reported in “Source Data” file.

    19. Source Data

      AssayResult: 90.64

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 1.82

      StandardErrorMean: 1.29

      Comment: Exact values reported in “Source Data” file.

    20. Source Data

      AssayResult: 11.37

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 0.36

      StandardErrorMean: 0.26

      Comment: Exact values reported in “Source Data” file.

    21. Source Data

      AssayResult: 70.86

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 17.18

      StandardErrorMean: 12.15

      Comment: Exact values reported in “Source Data” file.

    22. Source Data

      AssayResult: 81.81

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 7.45

      StandardErrorMean: 5.27

      Comment: Exact values reported in “Source Data” file.

    23. Source Data

      AssayResult: 90.54

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 10.24

      StandardErrorMean: 7.24

      Comment: Exact values reported in “Source Data” file.

    24. Source Data

      AssayResult: 13.45

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 2.2

      StandardErrorMean: 1.55

      Comment: Exact values reported in “Source Data” file.

    25. Source Data

      AssayResult: 92.2

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 19.94

      StandardErrorMean: 14.1

      Comment: Exact values reported in “Source Data” file.

    26. Source Data

      AssayResult: 90.79

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 6.38

      StandardErrorMean: 4.51

      Comment: Exact values reported in “Source Data” file.

    27. Source Data

      AssayResult: 69.83

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 5.94

      StandardErrorMean: 4.2

      Comment: Exact values reported in “Source Data” file.

    28. Source Data

      AssayResult: 75.85

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 4.78

      StandardErrorMean: 3.38

      Comment: Exact values reported in “Source Data” file.

    29. Source Data

      AssayResult: 94.33

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 9.99

      StandardErrorMean: 7.07

      Comment: Exact values reported in “Source Data” file.

    30. Source Data

      AssayResult: 102.58

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 2.19

      StandardErrorMean: 1.55

      Comment: Exact values reported in “Source Data” file.

    31. Source Data

      AssayResult: 21.79

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 1.84

      StandardErrorMean: 1.3

      Comment: Exact values reported in “Source Data” file.

    32. Source Data

      AssayResult: 95.86

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 1.62

      StandardErrorMean: 1.15

      Comment: Exact values reported in “Source Data” file.

    33. Source Data

      AssayResult: 91.21

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 7.32

      StandardErrorMean: 5.18

      Comment: Exact values reported in “Source Data” file.

    34. Source Data

      AssayResult: 10.59

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 0.6

      StandardErrorMean: 0.43

      Comment: Exact values reported in “Source Data” file.

    35. Source Data

      AssayResult: 76.97

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 3.1

      StandardErrorMean: 2.19

      Comment: Exact values reported in “Source Data” file.

    36. Source Data

      AssayResult: 7.92

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 1.32

      StandardErrorMean: 0.94

      Comment: Exact values reported in “Source Data” file.

    37. Source Data

      AssayResult: 38.85

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 3.05

      StandardErrorMean: 2.15

      Comment: Exact values reported in “Source Data” file.

    38. Source Data

      AssayResult: 16.89

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 3.12

      StandardErrorMean: 2.21

      Comment: Exact values reported in “Source Data” file.

    39. Source Data

      AssayResult: 17.62

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 1.76

      StandardErrorMean: 1.25

      Comment: Exact values reported in “Source Data” file.

    40. Source Data

      AssayResult: 86.41

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 17.62

      StandardErrorMean: 12.46

      Comment: Exact values reported in “Source Data” file.

    41. Source Data

      AssayResult: 6.16

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 1.27

      StandardErrorMean: 0.9

      Comment: Exact values reported in “Source Data” file.

    42. Source Data

      AssayResult: 95.16

      AssayResultAssertion: Not reported

      ReplicateCount: 3

      StandardDeviation: 16.94

      StandardErrorMean: 9.78

      Comment: Exact values reported in “Source Data” file.

    43. Source Data

      AssayResult: 7.08

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 0.54

      StandardErrorMean: 0.38

      Comment: Exact values reported in “Source Data” file.

    44. Source Data

      AssayResult: 14.01

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 0.53

      StandardErrorMean: 0.38

      Comment: Exact values reported in “Source Data” file.

    45. Source Data

      AssayResult: 74.49

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 2.29

      StandardErrorMean: 1.62

      Comment: Exact values reported in “Source Data” file.

    46. Source Data

      AssayResult: 6.53

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 0.29

      StandardErrorMean: 0.21

      Comment: Exact values reported in “Source Data” file.

    47. Source Data

      AssayResult: 30.27

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 1.04

      StandardErrorMean: 0.74

      Comment: Exact values reported in “Source Data” file.

    48. Source Data

      AssayResult: 7.63

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 0.51

      StandardErrorMean: 0.36

      Comment: Exact values reported in “Source Data” file.

    49. Source Data

      AssayResult: 63.4

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 5.92

      StandardErrorMean: 4.18

      Comment: Exact values reported in “Source Data” file.

    50. Source Data

      AssayResult: 60.28

      AssayResultAssertion: Not reported

      ReplicateCount: 3

      StandardDeviation: 0.14

      StandardErrorMean: 0.1

      Comment: Exact values reported in “Source Data” file.

    51. Source Data

      AssayResult: 17.35

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 6.21

      StandardErrorMean: 4.39

      Comment: Exact values reported in “Source Data” file.

    52. Source Data

      AssayResult: 106.23

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 14.57

      StandardErrorMean: 10.3

      Comment: Exact values reported in “Source Data” file.

    53. Source Data

      AssayResult: 75.71

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 22.31

      StandardErrorMean: 15.77

      Comment: Exact values reported in “Source Data” file.

    54. Source Data

      AssayResult: 6.66

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 0.28

      StandardErrorMean: 0.2

      Comment: Exact values reported in “Source Data” file.

    55. Source Data

      AssayResult: 6.1

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 2.11

      StandardErrorMean: 1.49

      Comment: Exact values reported in “Source Data” file.

    56. Source Data

      AssayResult: 84.07

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 2.47

      StandardErrorMean: 1.75

      Comment: Exact values reported in “Source Data” file.

    57. Source Data

      AssayResult: 100.07

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 6.18

      StandardErrorMean: 4.37

      Comment: Exact values reported in “Source Data” file.

    58. Source Data

      AssayResult: 91.6

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 8.2

      StandardErrorMean: 5.8

      Comment: Exact values reported in “Source Data” file.

    59. Source Data

      AssayResult: 82.83

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 4.82

      StandardErrorMean: 3.41

      Comment: Exact values reported in “Source Data” file.

    60. Source Data

      AssayResult: 87.35

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 11.94

      StandardErrorMean: 8.44

      Comment: Exact values reported in “Source Data” file.

    61. Source Data

      AssayResult: 83.25

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 5.27

      StandardErrorMean: 3.73

      Comment: Exact values reported in “Source Data” file.

    62. Source Data

      AssayResult: 7.03

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 2.68

      StandardErrorMean: 1.9

      Comment: Exact values reported in “Source Data” file.

    63. Source Data

      AssayResult: 77.45

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 6.2

      StandardErrorMean: 4.38

      Comment: Exact values reported in “Source Data” file.

    64. Source Data

      AssayResult: 9.92

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 1.93

      StandardErrorMean: 1.37

      Comment: Exact values reported in “Source Data” file.

    65. Source Data

      AssayResult: 95.02

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 0.08

      StandardErrorMean: 0.06

      Comment: Exact values reported in “Source Data” file.

    66. Source Data

      AssayResult: 10.4

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 3.22

      StandardErrorMean: 2.28

      Comment: Exact values reported in “Source Data” file.

    67. Source Data

      AssayResult: 7.75

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 2.59

      StandardErrorMean: 1.83

      Comment: Exact values reported in “Source Data” file.

    68. Source Data

      AssayResult: 75.45

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 4.03

      StandardErrorMean: 2.85

      Comment: Exact values reported in “Source Data” file.

    69. Source Data

      AssayResult: 98.55

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 5.74

      StandardErrorMean: 4.06

      Comment: Exact values reported in “Source Data” file.

    70. Source Data

      AssayResult: 62.31

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 11.49

      StandardErrorMean: 8.13

      Comment: Exact values reported in “Source Data” file.

    71. Source Data

      AssayResult: 66.19

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 21.26

      StandardErrorMean: 15.03

      Comment: Exact values reported in “Source Data” file.

    72. Source Data

      AssayResult: 105.41

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 9.45

      StandardErrorMean: 6.68

      Comment: Exact values reported in “Source Data” file.

    73. Source Data

      AssayResult: 7.82

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 2.31

      StandardErrorMean: 1.64

      Comment: Exact values reported in “Source Data” file.

    74. Source Data

      AssayResult: 92.32

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 2.26

      StandardErrorMean: 1.6

      Comment: Exact values reported in “Source Data” file.

    75. Source Data

      AssayResult: 44.9

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 9.75

      StandardErrorMean: 6.89

      Comment: Exact values reported in “Source Data” file.

    76. Source Data

      AssayResult: 97.61

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 0.97

      StandardErrorMean: 0.68

      Comment: Exact values reported in “Source Data” file.

    77. Source Data

      AssayResult: 11.28

      AssayResultAssertion: Abnormal

      ReplicateCount: 2

      StandardDeviation: 1.24

      StandardErrorMean: 0.87

      Comment: Exact values reported in “Source Data” file.

    78. Source Data

      AssayResult: 86.67

      AssayResultAssertion: Not reported

      ReplicateCount: 2

      StandardDeviation: 2.24

      StandardErrorMean: 1.58

      Comment: Exact values reported in “Source Data” file.

    79. We, therefore, analyzed the effect of 48 PALB2 VUS (Fig. 2a, blue) and one synthetic missense variant (p.A1025R) (Fig. 2a, purple)29 on PALB2 function in HR.

      HGVS: NM_024675.3:c.1010T>C p.(L337S)

    Tags

    Annotators

    URL

    1. Automated Patch ClampingCells were patch clamped with the SyncroPatch 384PE automated patch clamping device (Nanion). To prepare cells for patch clamping, cells were washed in PBS, treated with Accutase (Millipore-Sigma) for 3 min at 37°C, then recovered in CHO-S-serum free media (GIBCO). Cells were pelleted and resuspended in divalent-free reference solution (DVF) at ∼200,000–400,000 cells/mL. DVF contained (mM) NaCl 140, KCl 4, alpha-D(+)-glucose 5, HEPES 10 (pH 7.4) adjusted with NaOH. Cells were then added to a medium resistance (4–6 MΩ) 384-well recording chamber with 1 patch aperture per well (NPC-384, Nanion), which contained DVF and internal solution: CsCl 10, NaCl 10, CsF 110, EGTA 10, HEPES 10 (pH 7.2) adjusted with CsOH. Next, to enhance seal resistance, 50% of the DVF was exchanged with a calcium-containing seal enhancing solution: NaCl 80, NMDG 60, KCl 4, MgCl2 1, CaCl2 10, alpha-D(+)-glucose 5, HEPES 10 (pH 7.4) adjusted with HCl. The cells were washed three times in external recording solution: NaCl 80, NMDG 60, KCl 4, MgCl2 1, CaCl2 2, alpha-D(+)-glucose 5, HEPES 10 (pH 7.4) adjusted with HCl. Currents elicited in response to activation, inactivation, and recovery from inactivation protocols were then recorded (Figure S2). A late current measurement was captured every 5 s. After 1 min, 50% of the external solution was exchanged with external solution containing 200 μM tetracaine hydrochloride (Sigma; effective concentration 100 μM tetracaine). After tetracaine addition, late current measurements were obtained every 5 s for 1 additional minute. At least 10 cells expressing wild-type SCN5A were included for comparison in each SyncroPatch experiment (Figure 1), and at least 2 independent transfections and at least 10 replicate cells were studied per mutant. Recordings were performed at room temperature.We also conducted experiments to assess the effects of incubation at low temperature or mexiletine (a sodium channel blocker), interventions reported to increase cell surface expression of mistrafficked channels.27Clatot J. Ziyadeh-Isleem A. Maugenre S. Denjoy I. Liu H. Dilanian G. Hatem S.N. Deschênes I. Coulombe A. Guicheney P. Neyroud N. Dominant-negative effect of SCN5A N-terminal mutations through the interaction of Na(v)1.5 α-subunits.Cardiovasc. Res. 2012; 96: 53-63Crossref PubMed Scopus (62) Google Scholar,  28Makiyama T. Akao M. Tsuji K. Doi T. Ohno S. Takenaka K. Kobori A. Ninomiya T. Yoshida H. Takano M. et al.High risk for bradyarrhythmic complications in patients with Brugada syndrome caused by SCN5A gene mutations.J. Am. Coll. Cardiol. 2005; 46: 2100-2106Crossref PubMed Scopus (99) Google Scholar,  29Pfahnl A.E. Viswanathan P.C. Weiss R. Shang L.L. Sanyal S. Shusterman V. Kornblit C. London B. Dudley Jr., S.C. A sodium channel pore mutation causing Brugada syndrome.Heart Rhythm. 2007; 4: 46-53Abstract Full Text Full Text PDF PubMed Scopus (49) Google Scholar,  30Valdivia C.R. Ackerman M.J. Tester D.J. Wada T. McCormack J. Ye B. Makielski J.C. A novel SCN5A arrhythmia mutation, M1766L, with expression defect rescued by mexiletine.Cardiovasc. Res. 2002; 55: 279-289Crossref PubMed Scopus (77) Google Scholar,  31Valdivia C.R. Tester D.J. Rok B.A. Porter C.B. Munger T.M. Jahangir A. Makielski J.C. Ackerman M.J. A trafficking defective, Brugada syndrome-causing SCN5A mutation rescued by drugs.Cardiovasc. Res. 2004; 62: 53-62Crossref PubMed Scopus (106) Google Scholar For these experiments, cells stably expressing loss-of-function variants were generated as described above. The cells were incubated for 24 h at 30°C, or at 37°C with or without 500 μM mexiletine hydrochloride (Sigma), washed with HEK media, and were patch clamped as described above.

      AssayGeneralClass: BAO:0000062 patch clamp

      AssayMaterialUsed: CLO:0037237 HEK293-derived cell

      AssayDescription: HEK293T-derived cells stably expressing wild type or variant SCN5A were patch clamped and currents elicited in response to activation, inactivation, and recovery from inactivation were recorded, as well as late current measurements.

      AssayReadOutDescription: Peak current density relative to wild type, which was set to 100%

      AssayRange: %

      AssayNormalRange: Peak current density 75-125% of wild type

      AssayAbnormalRange: Peak current density 10-50% of wildtype

      AssayIndeterminateRange: Peak current density 50-75% of wildtype

      ValidationControlPathogenic: 0

      ValidationControlBenign: 10

      Replication: At least 2 independent transfections and at least 10 replicate cells per variant (see ReplicateCount in FunctionalAssayResult annotations for each variant).

      StatisticalAnalysisDescription: Two-tailed t tests or two-tailed Mann-Whitney U tests were used to compare variant parameters between groups of variants, while differences in dispersion between groups were tested with Levene’s test.

    2. Most Suspected Brugada Syndrome Variants Had (Partial) Loss of Function

      AssayResult: 100

      AssayResultAssertion: Normal

      ReplicateCount: 471

      StandardErrorMean: 3.7

      ControlType: Normal; wild type

      Comment: This variant (wildtype) had normal function. All other variant parameters were normalized to the values of wildtype. (Personal communication: A. Glazer)

    3. Most Suspected Brugada Syndrome Variants Had (Partial) Loss of Function

      AssayResult: 59.3

      AssayResultAssertion: Indeterminate

      ReplicateCount: 30

      StandardErrorMean: 8.3

      Comment: This variant had mild loss of function (peak current >50% and <75% of wildtype), therefore it was considered inconclusive and neither abnormal nor normal in vitro function. (Personal communication: A. Glazer)

    4. Most Suspected Brugada Syndrome Variants Had (Partial) Loss of Function

      AssayResult: 28.4

      AssayResultAssertion: Abnormal

      ReplicateCount: 13

      StandardErrorMean: 8.6

      Comment: This variant had partial loss of function of peak current (10-50% of wildtype), therefore it was considered abnormal (in vitro features consistent with Brugada Syndrome Type 1). (Personal communication: A. Glazer)

    5. Most Suspected Brugada Syndrome Variants Had (Partial) Loss of Function

      AssayResult: 45.3

      AssayResultAssertion: Abnormal

      ReplicateCount: 31

      StandardErrorMean: 5.1

      Comment: This variant had partial loss of function of peak current (10-50% of wildtype), therefore it was considered abnormal (in vitro features consistent with Brugada Syndrome Type 1).

    6. Most Suspected Brugada Syndrome Variants Had (Partial) Loss of Function

      AssayResult: 37.2

      AssayResultAssertion: Abnormal

      ReplicateCount: 26

      StandardErrorMean: 3.8

      Comment: This variant had partial loss of function of peak current (10-50% of wildtype), therefore it was considered abnormal (in vitro features consistent with Brugada Syndrome Type 1). (Personal communication: A. Glazer)

    7. Most Suspected Brugada Syndrome Variants Had (Partial) Loss of Function

      AssayResult: 23.1

      AssayResultAssertion: Abnormal

      ReplicateCount: 33

      StandardErrorMean: 3.2

      Comment: This variant had partial loss of function of peak current (10-50% of wildtype), therefore it was considered abnormal (in vitro features consistent with Brugada Syndrome Type 1). (Personal communication: A. Glazer)

    8. Most Suspected Brugada Syndrome Variants Had (Partial) Loss of Function

      AssayResult: 89.5

      AssayResultAssertion: Normal

      ReplicateCount: 29

      StandardErrorMean: 14.6

      Comment: This variant had normal function (75-125% of wildtype peak current, <1% late current, no large perturbations to other parameters). These in vitro features are consistent with non-disease causing variants. (Personal communication: A. Glazer)

    9. Most Suspected Brugada Syndrome Variants Had (Partial) Loss of Function

      AssayResult: 0.9

      AssayResultAssertion: Abnormal

      ReplicateCount: 18

      StandardErrorMean: 0.5

      Comment: This variant had loss of function of peak current (<10% of wildtype), therefore it was considered abnormal (in vitro features consistent with Brugada Syndrome Type 1). (Personal communication: A. Glazer)

    10. Most Suspected Brugada Syndrome Variants Had (Partial) Loss of Function

      AssayResult: 5.4

      AssayResultAssertion: Abnormal

      ReplicateCount: 19

      StandardErrorMean: 1.5

      Comment: This variant had loss of function of peak current (<10% of wildtype), therefore it was considered abnormal (in vitro features consistent with Brugada Syndrome Type 1). (Personal communication: A. Glazer)

    11. Most Suspected Brugada Syndrome Variants Had (Partial) Loss of Function

      AssayResult: 14.8

      AssayResultAssertion: Abnormal

      ReplicateCount: 27

      StandardErrorMean: 2.5

      Comment: This variant had partial loss of function of peak current (10-50% of wildtype), therefore it was considered abnormal (in vitro features consistent with Brugada Syndrome Type 1). (Personal communication: A. Glazer)

    12. Most Suspected Brugada Syndrome Variants Had (Partial) Loss of Function

      AssayResult: 78.9

      AssayResultAssertion: Normal

      ReplicateCount: 38

      StandardErrorMean: 7.2

      Comment: This variant had normal function (75-125% of wildtype peak current, <1% late current, no large perturbations to other parameters). These in vitro features are consistent with non-disease causing variants. (Personal communication: A. Glazer)

    13. Most Suspected Brugada Syndrome Variants Had (Partial) Loss of Function

      AssayResult: 43.3

      AssayResultAssertion: Abnormal

      ReplicateCount: 14

      StandardErrorMean: 12.2

      Comment: This variant had partial loss of function of peak current (10-50% of wildtype), therefore it was considered abnormal (in vitro features consistent with Brugada Syndrome Type 1). (Personal communication: A. Glazer)

    14. Most Suspected Brugada Syndrome Variants Had (Partial) Loss of Function

      AssayResult: 59.7

      AssayResultAssertion: Indeterminate

      ReplicateCount: 41

      StandardErrorMean: 6.3

      Comment: This variant had mild loss of function (peak current >50% and <75% of wildtype), therefore it was considered inconclusive and neither abnormal nor normal in vitro function. (Personal communication: A. Glazer)